This invention relates to the treatment or prophylaxis of conditions resulting from acidic gut syndrome in humans or an animals, wherein acidic gut syndrome results from the fermentation of carbohydrate in the gastrointestinal tract of the human or animal. The problems associated with acidic gut syndrome, and overcome with treatment, include: predisposition to ulceration of the gastrointestinal tract; ulceration of the stomach; immune conditions associated with localised inflammation of the gut including irritable bowel disorder, crohn""s disease, appendicitis, colitis and reduced feed intake, responsible for cachexia and low efficiency in production feeding systems; dennatitis; arthritis; rheumatoid arthritis; osteoarthritis; respiratory tract disorders, including asthma and predisposition to bleeding in lungs following strenuous exercise; predisposition to microbial and helminth infections of the gut, and infection of the mammary gland, including mastitis; immune disorders causing predisposition to infection by bacteria, fungi or protozoa; cystic fibrosis and certain cancers; effects on the pancreas, kidneys, thyroid and other organs and conditions of the endocrine system, including diabetes; homeostasis disorders, including blood pH, mineral and electrolyte imbalances, such as osteoporosis and hypertension; immune disorders, including multiple sclerosis, amyotrophic lateral sclerosis, chronic fatigue syndrome, myasthenia gravis, Alzheimer""s disease, impaired reproductive performance; dental caries; viral infections, including herpes; exacerbation of heat stress; and impaired hair and wool growth.
Carbohydrates are digested through enzymic breakdown and absorption of simple sugars or through microbial fermentation and absorption of short chain volatile fatty acids. A condition known as lactic acidosis (D-lactic acidosis, fermentative acidosis or carbohydrate overload) is widely recognised in ruminants and horses. This condition is responsible for deaths in ruminant livestock feeding, and in horses is associated with the development of laminitis (Garner et al. 1987, Rowe et al. 1994) and abnormal behaviour (Johnson et al. 1998). Lactic acidosis can also lead to diarrhoea, infections in the hind gut and skin disorders. The pathogenesis of lactic acidosis is described as a good example of metabolic acidosis in which considerable amounts of lactic acid are absorbed through the wall of the gut (Blood et al. 1983). D-lactic acid is more slowly metabolised than L-lactic acid and therefore accumulates in the tissues where it causes severe D-lactic acidosis. These authors also suggest that endotoxins from gram-negative bacteria in the gut may play a role in the pathogenesis of lactic acidosis (Blood et al. 1983), as it is possible that these endotoxins may be absorbed as a result of severe structural damage to the gut epithelium which occurs during this condition (Krueger et al. 1986).
The present invention describes a new condition, described as acidic gut syndrome, which differs significantly from lactic acidosis in that it does not involve metabolic acidosis, resulting from acid absorption from the gastrointestinal tract, as a factor in its pathogenesis. Acidic gut syndrome depends entirely on acidity within the gut and the adverse toxic effects mediated through the direct effect of acid on the gut wall and through effects of acidity on microbes within the gut. Acidic gut syndrome has a range of secondary consequences which develop from the primary effects of acidity within the gut. These secondary effects occur both locally and systemically. Locally, the effects are through the action of acidity and bacterial endotoxins (produced for example, through the death of gram negative bacteria under acidic conditions) on the gut wall itself. Systemically the effects are considered to be mediated via the immune system and a range of cytokine and related factors. Furthermore, it is thought that the gut wall and gut associated lymphoid tissue play a role in acidic gut syndrome through mediating the effects of toxins or other factors, and releasing systemically active hormones and/or agents of the immune system.
There are a number of immune system diseases, of a chronic and/or acute nature, which are of unknown aetiology. The common factor appears to be the involvement of various cytokines and other mediators of the immune system. Because the aetiology of these immune-related conditions has not been understood they have generally been considered to be non-specific immune diseases. Examples of immune diseases considered in this category are forms of arthritis, including rheumatoid arthritis, forms of respiratory disease and susceptibility to respiratory problems such as asthma, reduced enzyme production by the pancreas leading to some forms of diabetes, damage to kidneys resulting in mineral imbalances and hypertension, effects on the brain which can lead to secondary hormone imbalances with respect to control of temperature regulation, reproductive functions and other key aspects of metabolic control. The effects mediated via the immune system (cytokine and possibly other activities) can also cause inflammation and damage membranes in organs and tissues remote from the sites of bacterial activity within the gut. Organs and tissues affected in this way can include the lung (which subsequently increases the risk of respiratory tract infection), the stomach (which increases the risk of ulcer development), the kidneys (affecting mineral retention and hypertension). There can also be local areas of non-specific inflammation such as can occur in the gut or around the teeth. These conditions are considered to result from a minor build up of acidity within the gut and the release of endotoxins associated with the death of gram negative bacteria as acidity increases and gram positive bacteria predominate.
The slight increases in levels of both acidity and endotoxin within the gut associated with acidic gut syndrome, effect the host via a subtle increase in the immune challenge. The barrier between the host and the gastrointestinal microflora is extremely important in preventing infection and/or toxaemia. The present invention describes how the immune challenge to the host from the gut is not constant and can be increased in response to levels of acid in the gut which have until now been considered within normal limits and of no biological consequence. Increased acidity and changes in gut bacteria, particularly in relation to endotoxin production, produces a challenge which may not necessarily lead to an immediately detectable disease condition or a dramatic immunological response in the animal. It does however pose a sufficient challenge to the immune system of the animal or human to cause measurable responses (for example raised levels of tumour necrosis factor (TNF), T-cells, monocytes, interferon and cytokines, including interleukin-1, -6 and -8). Because these changes to the immune system are small and in themselves do not cause symptoms or signs of disease they have not previously been linked to changes in the diet or digestive process. Slightly elevated levels of cytokines and or TNF tend to be transitory and have previously been regarded as normal variation between individuals or non-specific immune conditions, and for these reasons have not been studied systematically with a view to determining the importance of diet and microbial activity within the gut. There has therefore been no previous suggestion that diet, fermentation within the gut, and subsequent acid accumulation and endotoxin production in the gut (acidic gut syndrome), are the primary causes of serious chronic and/or sporadic disease conditions of previously unknown origin.
There is a dense and diverse population of bacteria which inhabit the gastro-intestinal tract of man and animals. The concentration of these bacteria occurs naturally in those parts of the tract where the conditions of pH are close to neutral (around pH 6.5 to 7.5) and where fermentable substrate is available for fermentation. The bacteria in the gut poses a potential risk to the animal in terms of infection from the gut or through the absorption of microbial toxins.
The medical and nutritional literature contains information on various approaches used to manage the microbial population within the gut of humans. Two of these are summarised below.
1. The creation of a population of lactic acid producing bacteria in the gut through inoculation with probiotics such as cultures of Lactobacilli and other lactic acid producing bacteria, in the form of yoghurt cultures or specifically cultured bacterial preparations. The hypothesis behind this practice is that the lactic acid production may exclude other, more pathogenic, bacteria from the gut.
2. Consumption of increased amounts of soluble fibre which consists of indigestible (but fermentable) sources of starch (resistant starch) and oligosaccharides in order to provide substrates for fermentation in the hind gut. The aim of this practice is to increase the production of butyric acid, which has been shown, in vitro, to enhance the metabolism of the gut epithelium and reduce the risk of colonic cancer.
It is clear that these practices, now widely recommended by dietitians and medical practitioners, are likely to increase the amount of fermentable acid production and, specifically, the amount of lactic acid and are therefore likely to promote acidic gut syndrome.
The present invention describes how the elevated activity of the immune system, as a result of acidic gut syndrome, is sufficient to initiate, or predispose, the host to a range of secondary conditions including immune diseases, inflammation, infection and damage to membranes, by cytokine, or other immune system activity, and provides a method for the treatment or prophylaxis of acidic gut syndrome.
According to a first embodiment of this invention, there is provided a method for the treatment or prophylaxis of acidic gut syndrome resulting from the accumulation of acid and production of endotoxin in the gastrointestinal tract of a human or an animal, said accumulation resulting from the fermentation of carbohydrate in the gastrointestinal tract of said human or animal, wherein said method comprises administering to said human or animal an effective amount of an active agent capable of preventing or controlling acid and endotoxin accumulation in the gastrointestinal tract.
Typically, the active agent may be selected from the group consisting of: antibiotics, enzyme preparations, clay preparations, compounds which slow the digesta flow rate and probiotic preparations.
A suitable active agent of the first embodiment of the invention may include an antibiotic whose action is to control acid producing gram-positive bacteria.
Typically, the antibiotic may be selected from the group consisting of: a glycopeptide antibiotic, a glycolipid antibiotic, a staphylomycin antibiotic, a polypeptide antibiotic, a macrolide antibiotic, a sulphur-containing peptide antibiotic, a lincosamide antibiotic, tiamulin, a nitrofuran antibiotic, a tetracycline antibiotic, a penicillin antibiotic, a polythiazole antibiotic, an ionophore antibiotic, a cephalosporin antibiotic, a sulphonamide antibiotic, an aminoglycoside antibiotic, a quinalone antibiotic, streptogramin antibiotic, and any other antibiotic active against gram-positive bacteria responsible for the production of acid in the gastrointestinal tract.
Even more typically, the antibiotics active against gram-positive bacteria may be selected from the group consisting of: glycopeptide antibiotics, more typically, avoparcin, teicoplanin or vancomycin; glycolipid antibiotics, more typically flavomycin (bambemiycin); staphylomycin antibiotics, more typically virginiamycin; polypeptide antibiotics, more typically bacitracin zinc, bacitracin methylene disalicylate, virginiamycin S or polymixins (B and E); macrolide antibiotics, more typically tylosin, spiramycin, virginiamycin M, josamycin, spectinomycin or erythromycin; or sulfur-containing peptide antibiotics, more typically thiopeptone, thiopeptin, sulfomycin, thiostrepton, sporangiomycin, siomycin or taitomycin; lincosamide antibiotics, more typically lincomycin or clindamycin; or tiamulin; or nitrofuran antibiotics, more typically nitrofurantoin, nitrofurazone or furazolidone; tetracycline antibiotics, more typically chlortetracycline or oxytetracycline; penicillin antibiotics; more typically penicillinase-resistant penicillins, such as oxacillin or methicillin, penicillin V or ampicillin; polythiazole antibiotics, more typically nosiheptide; or ionophore antibiotics, more typically lasalocid, tetronasin, naracin or salinomycin; or ardacin, novobiocin sodium, bottromycin tartrate; streptogramin antibiotics, more typically, quinupristin/dalfopristin (RP 59500; Synercid) or streptogramin combinations [quinupristin/dalfopristin (RP 59500; Synercid)], everninomycin derivatives (SCH 27899), oxazolidinones (U-100572, U-100766); fluoroquinolone antibiotics, more typically, ciprofloxacin, ofloxacin, clinafloxacin, DU 6859a, grepafloxacin, levofloxacin, sparfloxacin or trovafloxacin; beta-lactam antibiotics; nitrovin (payzone), enramycin, mupiricin, margainin antibiotics, chloramphenicols and related compounds, including florphenicol, and any combination thereof.
A suitable active agent of the first embodiment of the invention may include an exogenous enzyme preparation designed to reduce the passage of fermentable carbohydrate to the hind gut through improving the digestion and absorption in the intestine of starches, disaccharides, oligosaccharides, non-starch polysaccharides, protein starch complexes and any polysaccharide which is incompletely digested in the intestine, but which is readily fermentable in the hind gut.
Typically preferred enzymes for the break down of non-starch polysaccharides and starches include the following: glyconases including: amylase, maltase, invertase, xcex1-glucosidases, emulsin, and amyloglucosidase; glucanasess, xcex2-glucanase, xylanase; enzymes which break down galactosides of the raffinosse series and other xcex1-galactosides including xcex1-galactosidase, enzymes which break down the proteins forming part of the matrix surrounding starches, sugars and non-starch carbohydrates in plant material, including: pepsin, trypsin, trypsinogen, chymotrypsin and natural and synthetic proteolytic enzymes of chemical or microbial origin, enzymes which depolymerise non-starch polysaccharides including: arabinoxylans and xcex1-glucans, and enzymes active in the break down of cellulose, including: cellulase, enzymes active in the break down of colloidal polysaccharides, pectic substances, which include: galactouronans, galactan and arabinans, as well as the neutral polysaccharides such as xyloglucans and galactomannans and other non-starch polysaccharides such as: rhamnogalactouronan with arabinose and galactose, arabinogalactan, glucan, xyloglucan, galactouronan with arabinose and uronan with arabinose. These enzymes can be used individually or in combination.
A suitable active agent of the first embodiment of the invention may include a clay preparation which reduces the rate of fermentation and binds specific ions in a way which reduces the adverse effects of rapid fermentation of starch and other soluble carbohydrates in the gastrointestinal tract.
Typically, preferred clays for reducing the rate of fermentation and the osmotic effects of rapid fermentation within the gut include: kaolinite, bentonite, montmorrilonite, illite, clinoliptolite, heulandite, palygorsite, saponite, smectite, chrysotile, lizardite, talc, pyrophyllite, vermiculite, beidellite, halloysite or zeolite types of clay, and these can be activated by a wide range of ions including sodium, calcium, potassium and mixtures of these and other ions. These clays can be used individually or in combination.
A suitable active agent of the first embodiment of the invention may include a compound which slows digesta flow rate, thereby increasing intestinal digestion and absorption and reducing the amount of fermentable substrate passing to the hind gut.
Generally, preferred agents to slow the flow of digeseta include biologically active peptides (BAP) in a form which will reach the duodenum, and are active in modulating the activity of the digestive tract, gastric emptying and the rate of passage through the intestine. More typically, these biologically active peptides include opioid peptides.
Whilst a range of proteins potentially produce opioid peptides on hydrolisation, the xcex2-casomorphins, which can be derived from xcex2-casein during xcex2-casein digestion, are particularly active.
Even more typically, the biologically active peptides include cholecystokinin (CCK), the M1 fraction of virginiamycin and the analogue of virginiamycin fraction M1, compound L-156. These biologically active peptides can be used individually or in combination.
It has traditionally been assumed that the nutritional benefits of proteins are only related to the essential amino acids supplied to the animal during digestion and absorption. However through the supply of biologically active peptides and the production of naturally occurring opioid peptides, the rate of digesta passage is reduced and this results in more efficient intestinal digestion and less fermentable substrate passing to the hind gut which can contribute to acidic gut syndrome.
Practical methods of supplying biologically active opioid peptides is through dietary supplementation with proteins such as casein and blood meal. For ruminant animals the best results are obtained through protection of the protein against rumen degradation by polymer coating technology, slow-release capsules, or through formaldehyde treatment.
A suitable active agent of the first embodiment of the invention may include a probiotic preparation which reduces lactic acid accumulation by: formation of alternative end products of fermentation; production of volatile fatty acids rather than lactic acid during carbohydrate fermentation; through increased utilisation of lactic acid; or through the conversion of lactic acid to volatile fatty acids which can be absorbed from the gut, thereby reducing acidity in the gut.
Typically, preferred probiotic preparations include bacteria which ferment starch and sugars to end products other than lactic acid, (ie volatile fatty acids).
More typically, the probiotic preparations may include bacteria selected from the group consisting of: Succinonionas, Butyrivibrio, Bacteroides and Succinivibrio. These bacteria can be used individually or in combination.
Typically, preferred probiotic preparations include bacteria capable of utilising lactic acid, and converting lactic acid to volatile fatty acids and other end products.
More typically, the probiotic preparations may include anaerobic bacteria.
Even more typically, the probiotic preparations may include bacteria selected from the group consisting of: Megasphera, Veillenolla, Selenomonas, Propionibacterium, Anaerovibrio and Peptococcus. These bacteria can be used individually or in combination.
Typically, preferred probiotic preparations include yeast and mycelial preparations capable of utilising lactic acid, and converting lactic acid to volatile fatty acids and other end products.
More typically, the probiotic preparations may include yeast and mycelial preparations such as Yea Sacc.
Typically, at least any two of the above sample microrganisms of the probiotic preparation may be used in combination in the probiotic preparation.
According to a further aspect of the invention, the active agent may include a combination of at least two of: an antibiotic, an exogenous enzyme preparation, a clay, a compound which slows digesta flow rate, or a probiotic preparation.
Compositions for administration of the active agent in the method of the invention may be prepared by means known in the art for the preparation of compositions (such as in the art of veterinary and pharmaceutical compositions) including blending, grinding, homogenising, suspending, dissolving, emulsifyng, dispersing and where appropriate, mixing of the active agent, together with selected excipients, diluents, carriers and adjuvants.
For oral administration, the pharmaceutical or veterinary composition may be in the form of tablets, lozenges, pills, troches, capsules, elixirs, powders, including lyophilised powders, solutions, granules, suspensions, emulsions, syrups and tinctures. Slow-release, or delayed-release, forms may also be prepared, for example in the form of coated particles, multi-layer tablets or microgranules.
Solid forms for oral administration may contain binders acceptable in human and veterinary pharmaceutical practice, sweeteners, disintegrating agents, diluents, flavourings, coating agents, preservatives, lubricants and/or time delay agents. Suitable binders include gum acacia, gelatine, corn starch, gum tragacanth, sodium alginate, carboxymethylcellulose or polyethylene glycol. Suitable sweeteners include sucrose, lactose, glucose, aspartame or saccharine. Suitable disintegrating agents include corn starch, methylcellulose. polyvinylpyrrolidone, guar gum, xanthan gum, bentonite, alginic acid or agar. Suitable diluents include lactose, sorbitol, mannitol, dextrose, kaolin, cellulose, calcium carbonate, calcium silicate or dicalcium phosphate. Suitable flavouring agents include peppermint oil. oil of wintergreen, cherry, orange or raspberry flavouring. Suitable coating agents include polymers or copolymers of acrylic acid and/or methacrylic acid and/or their esters, waxes, fatty alcohols, zein, shellac or gluten. Suitable preservatives include sodium benzoate, vitamin E, alpha-tocopherol, ascorbic acid, methyl paraben, propyl paraben or sodium bisulphite. Suitable lubricants include magnesium stearate, stearic acid, sodium oleate, sodium chloride or talc. Suitable time delay agents include glyceryl monostearate or glyceryl distearate.
Liquid forms for oral administration may contain, in addition to the above agents, a liquid carrier. Suitable liquid carriers include water, oils such as olive oil, peanut oil, sesame oil, sunflower oil, safflower oil, arachis oil, coconut oil, liquid paraffin, ethylene glycol, propylene glycol, polyethylene glycol, ethanol, propanol, isopropanol, glycerol, fatty alcohols, triglycerides or mixtures thereof.
Suspensions for oral administration may further include dispersing agents and/or suspending agents. Suitable suspending agents include sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethyl-cellulose, poly-vinyl-pyrrolidone, sodium alginate or acetyl alcohol. Suitable dispersing agents include lecithin, polyoxyethylene esters of fatty acids such as stearic acid, polyoxyethylene sorbitol mono- or di-oleate, -stearate or -laurate, polyoxyethylene sorbitan mono- or di-oleate, -stearate or -laurate and the like.
The emulsions for oral administration may further include one or more emulsifying agents. Suitable emulsifying agents include dispersing agents as exemplified above or natural gums such as guar gum, gum acacia or gum tragacanth.
In another form the invention resides in a method of treating animals or humans which comprises delivering to the alimentary canal a quantity of an active agent of the form described above.
Typically the active agent can be administered by binding it to fibrous materials which pass undigested into the caecum, colon or other part of the hind gut or it can be incorporated into specially formulated feeds and foods or administered in the form of premixes, pastes, gels, gums, pellets or cubes. In one particular form of the invention, administration of the active agent to human or animal subjects is in the form of digestible capsules which release the active material into the stomach, intestine or hindgut.
The active agents may be administered with the carbohydrate feed. The active agent can be mixed with the feed during preparation, added to the feed before consumption or oral administration or sprinkled on top of the food before it is consumed. In one particular form of the invention, in the administration of the active agent to human subjects, the active agent can be mixed with herbs and spices coating starch based foods such as biscuits and snack foods. Further, the active agent can be included in pelleted feeds for animals and/or in loose mixes.
Typically, the active agent may be administered directly into the buccal cavity.
Typically, administration of the active agent to human subjects may be in the form of toothpaste which releases the active agent into the buccal cavity.
Typically, the active agent may be administered to the human or animal subjects via targeted delivery to the hind gut using enteric coated delivery systems to ensure specific activity of the active agent in the terminal ileum, colon and/or caecum.
Typically, targeted delivery to different parts of the gastrointestinal system is achieved by multiple coatings of the active agent with materials sensitive to pH and/or enzyme activity, and/or microbial fermentation and/or time-dependent solubility.
Typically, a method of treating animals in accordance with the present invention may include delivering to the alimentary canal a quantity of between 1 and 10% of total feed material of an active agent of the form described above, and 90-99% feed material wherein the feed material includes between 1 and 10% of fibrous material. Typically the fibrous material may be lucerne chaff.
The administered dose of the antibiotic can vary and will depend on several factors, such as the condition, age and size of the human or animal patient, as well as the nature of the lactic acid producing gram-positive bacteria. Dosages will typically range from between 0.01 and 5 mg per kg of bodyweight. More typically dosages will range from between 0.02 and 2.0 mg per kg of bodyweight. More typically dosages will range from between 0.05 and 1.0 mg per kg of bodyweight. Even more typically dosages will range from between 0.1 and 0.5 mg per kg of bodyweight. Yet even more typically, the antibiotic is administered to the human or animal at a rate of 0.4 mg per kg of bodyweight.
Typically, the antibiotic is administered at a rate of between 1 and 100 mg per kg of dry weight of food. More typically, the antibiotic is administered at a rate of between 1 and 75 mg per kg of dry weight of food. Even more typically, the antibiotic is administered at a rate of between 1 and 50 mg per kg of dry weight of food. Yet even more typically, the antibiotic is administered at a rate of between 5 and 40 mg per kg of dry weight of food.
As above, the administered dose of the enzyme preparation can vary and will depend on several factors, such as the condition, age and size of the human or animal patient, as well as the nature of the carbohydrate. Dosages will typically range from between 0.01 and 50 g/kg food dry matter. Typically, the enzyme is administered at a rate of between 0.1 and 3 g per kg of dry weight of food. More typically, the enzyme is administered at a rate of between 1 g per kg of dry weight of food.
Similarly, the administered dose of the clay preparation can vary and will depend on several factors, such as the condition, age and size of the human or animal patient, as well as the nature of the carbohydrate. Dosages will typically range from between 0.5 and 100 g/kg food dry matter. Typically, the clay is administered at a rate of between 1 and 50 g per kg of dry weight of food. More typically, the clay is administered at a rate of between 10 and 20 g per kg of dry weight of food.
Typically, the administered dose of the probiotic preparation can vary between 106 and 1012 bacteria per kg of body weight. More typically, dose of the probiotic preparation can vary between 108 and 1010 per kg of body weight.
According to another form of the invention, the active agents can be used together.
According to another aspect of the invention, the formulation of the active agent ensures that it is administered in a palatable form to the animal or human and in a form which retains activity and is properly mixed in the appropriate compartment(s) of the gastrointestinal tract.
Generally, the active agent is administered regularly throughout the period the animal or human is subjected to a high carbohydrate diet or to sugars or other fermentable compounds which are not efficiently absorbed prior or reaching the large intestine, colon and caecum.
More typically, the active agent is administered 1-3 times daily. Even more typically, the active agent is administered once daily or can be included in human food and animal feeds. They can be fed as powders or suspended in water, included in pellets as well as being fed in premixes.
More typically the active agent is mixed with the food, or is added to feeds which contain starch or sugars which may produce an acidic pattern of fermentation in the gastrointestinal tract.
A suitable treatment may include the administration of a single dose or multiple doses. Usually, the treatment will consist of administering one dose daily of the active agent for a period sufficient to control the accumulation of acid by fermentation of the carbohydrate in the gastrointestinal tract. Dosing may continue while sources of carbohydrate known to cause problems of acidic fermentation in the gastrointestinal tract are included in the diet.
More typically the active agent may be administered in a single dose immediately before consuming meals containing sources of carbohydrate which are poorly digested and rapidly fermented.
More typically, the active agent is administered for one day prior to and daily during the consumption of excessive quantities of food stuffs containing readily fermentable carbohydrates.
Typically, the active agent is administered orally.
According to a second embodiment of the invention, there is provided a method for the treatment or prophylaxis of acidic gut syndrome resulting from the accumulation of acid and production of endotoxin in the gastrointestinal tract of a human or an animal, said accumulation resulting from the fermentation of carbohydrate in the gastrointestinal tract of said human or animal, which method comprises immunising said human or animal against the microorganisms responsible for the fermentation of carbohydrate in the gastrointestinal tract of said human or animal.
Typically, the human or animal is immunised against bacteria which produce lactic acid in the gut and therefore primary agents in the development of acidic gut syndrome.
Typically, bacteria against which the human or animal is immunised include: Aerococcus, Alloiococcis, Carnobacterium, Enterococcus, Lactobacillus, Lactococcus, Leuconostoc, Pediococcu, Streptococcu and Tetragenacoccus among others.
More typically, bacteria against which the human or animal is immunised include: Lactobacillus spp. and Streptococcus bovis type.
Even more typically, bacteria against which the human or animal is immunised is Streptococcus bovis (Sb-5).
For example, immunisation may be achieved by intramuscular or sub-cutaneous injection of for example, a mixture of Lactobacillus spp. and Streptococcus bovis, or either bacteria administered individually, together with a suitable adjuvant, excipient, diluent and/or carrier.
Typically, the adjuvant may include Quil A, Dex and Alum, cytokines, among others, and be of a variety of types suitable for different host species.
Typically, numerous strains of Lactobacillus spp. and Streptococcus bovis are suitable, and can be cultured using carbohydrate-rich media from rumen contents, caecal digesta or faeces.
Typically, a priming dose is followed by regular boosters to maintain immunity.
Typically, the dosage rate for immunisation is between 1xc3x97109 and 1xc3x971011 bacterial cells per injection.
Typically, the dosage rates are approximately equivalent to between 1xc3x97108 to 1xc3x97109 bacterial cells per kg body weight.
More typically, the dosage rates are approximately equivalent to between 1xc3x97108 and 5xc3x97108 bacterial cells per kg body weight.
Even more typically, the dosage rates arc approximately equivalent to 2.5xc3x97108 bacterial cells per kg body weight.
Typically, the dosage rate for immunisation of small animals, such as sheep, is between 5xc3x97109 and 5xc3x971010 bacterial cells per injection.
More typically, the dosage rate for immunisation of small animals, such as sheep, is approximately 1xc3x971010 bacterial cells per injection.
Typically, the dosage rate for immunisation of large animals, such as cattle and horses, is between 1xc3x971010 and 1xc3x971012 bacterial cells per injection.
More typically, the dosage rate for immunisation of large animals, such as cattle and horses, is approximately 1xc3x971011 bacterial cells per injection.
Typically, the injection volume for sheep is between 1 mL to 3 mL, and 2 to 7 mL for cattle and horses 3 to 5 mL.
More typically, the injection volume for sheep is between 1 mL to 2 mL, and 3 to 5 mL for cattle and horses.
The methods of the first or second embodiments of the invention are effective in the treatment or prophylaxis of the conditions associated with acidic gut syndrome.
The conditions associated with acidic gut syndrome, and treated by the methods of the first or second embodiments of the invention, include: predisposition to ulceration of the gastrointestinal tract; ulceration of the stomach; immune conditions associated with localised inflammation of the gut including irritable bowel disorder, crohn""s disease, appendicitis, colitis and reduced feed intake, responsible for cachexia and low efficiency in production feeding systems
The conditions associated with acidic gut syndrome, and treated by the methods of the first or second embodiments of the invention, even further include: dermatitis; arthritis; rheumatoid arthritis; osteoarthritis; respiratory tract disorders, including asthma and predisposition to bleeding in lungs following strenuous exercise.
The conditions associated with acidic gut syndrome, and treated by the methods of the first or second embodiments of the invention, still further include: predisposition to microbial and helminth infections of the gut, and infection of the mammary gland, including mastitis.
The conditions associated with acidic gut syndrome, and treated by the methods of the first or second embodiments of the invention, yet still further include: immune disorders causing predisposition to infection by bacteria, fungi or protozoa; cystic fibrosis and certain cancers.
The conditions associated with acidic gut syndrome, and treated by the methods of the first or second embodiments of the invention, yet still even further include: effects on the pancreas, kidneys, thyroid and other organs and conditions of the endocrine system, including diabetes; homeostasis disorders, including blood pH, mineral and electrolyte imbalances, such as osteoporosis and hypertension.
The conditions associated with acidic gut syndrome, and treated by the methods of the first or second embodiments of the invention, also include: immune disorders, including multiple sclerosis, amyotrophic lateral sclerosis, chronic fatigue syndrome, myasthenia gravis, Alzheimer""s disease, impaired reproductive performance; dental caries; viral infections, including herpes; exacerbation of heat stress; and impaired hair and wool growth.
According to a third embodiment of the invention, there is also provided a method of diagnosing acidic gut syndrome.
Typically, the method for diagnosing acidic gut syndrome comprises determining the pH of the gastrointestinal tract of said human or animal.
More typically, the pH of the gastrointestinal tract may be determined through pH indicator solutions or devices added to the toilet bowl, or pH indicator solutions or devices, used in conjunction with stool collections chambers.
Typically, the pH indicator solutions may involve pH sensitive colour reagents.
More typically, the pH indicator devices, used in conjunction with stool collections chambers, may involve paper strips.
Even more typically, the paper strips are litmus paper.
Yet even more typically, the litmus paper may be incorporated into toilet paper.
Typically, the acid which accumulates in the gastrointestinal tract includes: volatile fatty acids, and/or lactic acid.
Typically, the test for acidity may involve measurement of specific acids present in the faecal material, such as volatile fatty acids, and/or lactic acid.
Typically, acid measurement is performed using specific quantitative measurements, wherein such measurements may be made using chromatographic or spectroscopic techniques to measure individual acid concentrations.
Typically, the method for diagnosing acidic gut syndrome comprises determining the concentration of cytokines in the gastrointestinal tract, faeces and/or plasma of said human or animal.
Typically, the cytokine measured is selected from the group consisting of: tumour necrosis factor (TNF), interferon and interleukin.
Generally, the level of cytokine is measured via radioimmunoassay, ELISA or other molecular biological detection techniques.
Typically, the method for diagnosing acidic gut syndrome comprises determining the presence of bacterial endotoxins/lipopolysaccharides in the gastrointestinal tract of said human or animal.
The bacterial endotoxins/lipopolysaccharides may be detected through nucleic acid hybridisation and/or amplification.
Typically, hybridisation detection occurs via a nucleic acid probe specific for a bacterial endotoxin/lipopolysaccharide.
Typically, nucleic acid amplification may also provide a method of detection, and may occur via a pair of nucleic acid primers specific for a bacterial endotoxin/lipopolysaccharide.
More typically, amplification may be carried out using the ligase chain reaction (LCR).
Even more typically, amplification may be carried out using the polymerase chain reaction (PCR).
The bacterial endotoxins/lipopolysaccharides may be detected via an antibody detection system.
Typically, the detection system may involve the use of at least one polyclonal antibody.
More typically, the detection system may involve the use of at least one monoclonal antibody.
Typically, bacterial endotoxin/lipopolysaccharide may be measured using the limulus amoebocyte lysate system.